逆转录病毒介导的靶向人Slingshot-1L基因的稳定转染细胞系的建立

Establishment of stable transfected cell line Slingshot-1L mediated by retroviral

目的应用Slingshot(SSH)-1L重组逆转录病毒载体,建立过表达该基因的稳定转染细胞系。方法用脂质体法将含SSH-1L的重组逆转录病毒载体pLNCX-SSH-1L转染PA317包装细胞,G418筛选,滴度测定。收集病毒上清感染MG63细胞,G418筛选。结果应用pLNCX-Slingshot-1L重组逆转录病毒载体,获得滴度为5×108 CFU/L的病毒上清,感染MG63细胞,获得了过表达SSH-1L基因的MG63细胞。结论应用SSH-1L重组逆转录病毒载体,成功建立了过表达该基因的稳定细胞系。

Objective To establish stable transfected cell line by retroviral vector encoding Slingshot-1L.Methods The plasmid pLNCX-Slingshot-1L was transfected into packaging cell PA317 by lipofectamine. The transfectants were selected by G418 and viral titers were analyzed. Virals supernatant were collected to infect MG63 sells,selected by G418. Results Virals supernatant with high viral titers were obtained by retroviral vector encoding Slingshot-1L. Virals supernatant were collected to infect MG63 sells,and the stable transfected MG63 cell line was established. Conclusions The stable transfected cell line over-expression Slingshot-1L mediated by retroviral is established successfully.